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Measuring endosomal pH beyond the diffraction limit

Douglas Richardson

Harvard Center for Biological Imaging

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Fluorescence-based biosensors have become essential tools for modern biology, allowing real-time monitoring of biological processes within living cells. One widely used application of biosensors is to monitor the acidification of vesicles during endocytosis, an essential function that aids in intracellular cargo sorting and degradation. Prior to the development of super-resolution fluorescence microscopy (nanoscopy), investigation of endosomal dynamics in live cells remained difficult as these structures lie at or below the ~250鈥塶m diffraction limit of light microscopy. Therefore, to aid in investigations of pH dynamics during endocytosis at the nanoscale, we have specifically designed a family of ratiometric endosomal pH probes for use in live-cell STED nanoscopy.